Directed mutagenesis of the transmembrane domain of the PsbL subunit of photosystem II in Synechocystis sp. PCC 6803.
The PsbL protein is one of three low-molecular-weight subunits identified at the monomer-monomer interface of photosystem II (PSII) [Ferreira et al. (2004) Science 303:1831-1838; Loll et al. (2005) Nature 438:1040-1044]. We have employed site-directed mutagenesis to investigate the role of PsbL in Synechocystis sp. PCC 6803 cells. Truncation of the C-terminus by deleting the last four residues (Tyr-Phe-Phe-Asn) prevented association of PsbL with the CP43-less monomeric sub-complex and therefore blocked PSII assembly resulting in an obligate photoheterotrophic strain. Replacement of these residues with Ala created four photoautotrophic mutants. Compared to wild type, the F37A, F38A, and N39A strains had reduced levels of assembled PSII centers and F37A and F38A cells were readily photodamaged. In contrast, Y36A and Y36F mutants were similar to wild type. However, each of these strains had elevated levels of the CP43-less inactive monomeric complex. Mutations targeting a putative hydrogen bond between Arg-16 and sulfoquinovosyldiacylglycerol resulted in mutants that were also highly susceptible to photodamage. Similarly mutations targeting a conserved Tyr residue (Tyr-20) also destabilized PSII under high light and suggest that Tyr-20-lipid interactions or interactions of Tyr-20 with PsbT influence the ability of PSII to recover from photodamage.
Luo
Hao
H
Department of Biochemistry, University of Otago, Dunedin, New Zealand.
Eaton-Rye
Julian J
JJ
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010788:Photosynthesis; D045332:Photosystem II Protein Complex / Q000235:genetics / Q000502:physiology*; D016415:Sequence Alignment; D046939:Synechocystis / Q000235:genetics / Q000502:physiology*
D001426:Bacterial Proteins; D045332:Photosystem II Protein Complex; C075419:photosystem II, psbL subunit